CN103045713A - Method for detecting trace amounts of microbes in a liquid sample containing multiple impurities - Google Patents
Method for detecting trace amounts of microbes in a liquid sample containing multiple impurities Download PDFInfo
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- CN103045713A CN103045713A CN2013100189339A CN201310018933A CN103045713A CN 103045713 A CN103045713 A CN 103045713A CN 2013100189339 A CN2013100189339 A CN 2013100189339A CN 201310018933 A CN201310018933 A CN 201310018933A CN 103045713 A CN103045713 A CN 103045713A
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Abstract
The invention discloses a device and method for detecting trace amounts of microbes in a liquid sample containing multiple impurities. The method comprises the following steps: separating impurities and homogenizing the liquid with a membrane separation device and a homogenizer, carrying out vacuum filtration and enrichment with a microbial membrane filter, and finally, detecting after culturing. The device and method disclosed by the invention can be utilized to detect microbes in the massive liquid sample containing multiple solids and other large-granularity impurities as well as trace amounts of microbes, and have the advantages of high detection speed, favorable impurity removal accuracy and high efficiency.
Description
Technical field
The present invention relates to the detection method of a kind of microorganism, particularly the detection method of micro-microorganism in a kind of polymictic liquid sample.
Background technology
Other environment waters such as water of river and lake, process water, seawater contain a large amount of solid impurities usually, a small amount of microorganism when estimating its clean level or needing certain microorganism of other aimed detection, often is difficult to other impurity of fine separation, and complex operation produces error easily.
Summary of the invention
The objective of the invention is to overcome the deficiencies in the prior art, the apparatus and method of micro-microorganism in a kind of many impurity liquid sample are provided.
For achieving the above object, the present invention takes following technical proposals to realize:
Micro-microorganism detection method in a kind of polymictic liquid sample may further comprise the steps:
A. get a certain amount of liquid sample to be detected, put into membrane separation unit, then membrane separation unit is placed slap type homogenizer;
B., velocity of flapping and homogenizing time are set, separate the solid impurity in the testing sample, and so that the liquid sample in the membrane separation unit is full and uniform;
C. to each parts of microbial film filtration unit, comprise that stainless steel funnel and funnel web member sterilize, then connecting pipeline is closed each valve on the stainless steel stent, opens vacuum pump, regulate vacuum tightness, tweezers with after the sterilization are placed on aseptic filter membrane on the stainless steel filter membrane supporting pad, liquid sample behind the absorption separation homogeneous, pour in the stainless steel funnel to corresponding scale marks, open respective valves and carry out suction filtration;
D. after suction filtration is finished, touch the stainless steel funnel to a side, then upwards take out the stainless steel funnel, take off aseptic filter membrane with the antimagnetic type tweezer after the sterilization, carry out corresponding detection;
E. the aseptic filter membrane after the suction filtration enrichment is placed the sterilized culture dish that contains substratum, cultivate by inspecting standard, again detect after the cultivation.
Further, to the liquid sample of small amount, quantitative sample and diluent are added in the membrane separation unit simultaneously.
Preferably, to each parts of microbial film filtration unit, comprise that stainless steel funnel and funnel web member carry out flame sterilization.
The proofing unit of micro-microorganism comprises membrane separation unit in a kind of polymictic liquid sample, slap type homogenizer, microbial film filtration unit.
Wherein, described membrane separation unit comprises the bag that two sides form, the sealing of bag four limits, top is for can remove part, the bottom is the bag main body, and inside is provided with filter membrane, and the base of described filter membrane and top are individually fixed in base and the top of bag, dual-side is individually fixed on the two sides of bag, and the width of filter membrane is less than the width of bag side.
Preferably, the dual-side of described filter membrane is individually fixed in the mid-way of bag two sides.
Wherein, described microbial film filtration unit comprises stainless steel stent, at least one stainless steel filter, the stainless-steel vacuum table, vacuum filtration bottle, vacuum pump, wherein vacuum meter and stainless steel filter are installed on the stainless steel stent by web member, the vacuum filtration bottle is housed between stainless steel stent and the vacuum pump, and stainless steel stent is connected pipe connecting with the vacuum filtration bottle and connects, and described stainless steel filter comprises stainless steel base and the stainless steel funnel that is installed on the stainless steel base.
Preferably, described stainless steel funnel is connected on the stainless steel base by the mode of inlay card.
Preferably, described stainless steel base is provided with stainless steel filter membrane supporting pad.
The present invention has following technique effect: by adopting preceding method and device, isolate solid and other large size impurity in the liquid sample, then realize the full and uniform of sample, carry out the suction filtration enrichment by the microbial film filtration unit, detect through cultivating again at last, the convenient detection contained many kinds of solids and other large granular impurities, and contain the microorganism detection in a large amount of liquid samples of a small amount of microorganism, detection speed is fast, despumation, accuracy is good, and efficient is high;
Further, to the liquid sample of small amount, quantitative sample and diluent are added in the membrane separation unit simultaneously, after the dilution, can make things convenient for follow-up suction filtration operation, owing to carried out homogeneous, need not to worry the microbial profile heterogeneity simultaneously, cause the problem of resultant error;
Further, before carrying out suction filtration, the microbial film filtration unit is carried out flame sterilization, rapidly and efficiently, the while can cool off fast, can drop into rapidly to reuse, and saves detection time, improves detection efficiency.
Description of drawings
Fig. 1 is the schema of the detection method of micro-microorganism in a kind of polymictic liquid sample in the embodiment of the invention.
Fig. 2 is the structural representation of microbial film filtration unit in the embodiment of the invention.
Fig. 3 is the structural representation of membrane separation unit in the embodiment of the invention.
Fig. 4 is the expansion synoptic diagram of membrane separation unit in the embodiment of the invention.
Embodiment
The present invention will be described in detail below in conjunction with accompanying drawing.
Fig. 1 is the schema of the detection method of micro-microorganism in a kind of polymictic liquid sample in the embodiment of the invention.As shown in Figure 1, micro-microorganism detection method in this polymictic liquid sample may further comprise the steps:
A. draw sample 11, get a certain amount of liquid sample to be detected, put into membrane separation unit, then membrane separation unit is placed slap type homogenizer.
In the present embodiment, more such as the testing liquid sample size, then can directly put into membrane separation unit, if when the liquid sample amount is less, quantitative sample and diluent can be added in the membrane separation unit simultaneously, to reach the purpose of dilute sample.
The present invention can select the multiple film tripping device that is suitable for and homogenizing bag etc.Fig. 3 is the structural representation of the membrane separation unit that adopts among the better embodiment of the present invention, Fig. 4 is the expansion synoptic diagram of described membrane separation unit, can find out in conjunction with Fig. 3 and Fig. 4, described membrane separation unit comprises the bag that two sides form, the sealing of bag four limits, top is for can remove part 31, the bottom is bag main body 32, inside is provided with filter membrane 33, the base of described filter membrane 33 and top are individually fixed in base and the top of bag, dual-side is individually fixed on the mid-way of two sides of bag, and the width of filter membrane 33 is less than the width of bag side.
Described filter membrane 33 is divided into two spaces with membrane separation unit, during application of sample, testing sample is added in one of them space.
B. homogeneous separates 12, and velocity of flapping and homogenizing time are set, and separate the solid impurity in the testing sample, and so that the liquid sample in the membrane separation unit is full and uniform.
Difference per sample can arrange suitable velocity of flapping and time, and bounces spacing etc.Cooperation by slap type homogenizer, solid impurity in the liquid sample in the membrane separation unit and large-size particle are trapped within the space of the initial application of sample of filter membrane one side, and the sample in the space of filter membrane opposite side removes clear solid impurity substantially, and microbial profile is even in the sample, can proceed next step operation.
C. the suction filtration enrichment 13, to each parts of microbial film filtration unit, comprise that stainless steel funnel and funnel web member sterilize, then connecting pipeline, close each valve on the stainless steel stent, open vacuum pump, regulate vacuum tightness, with the tweezers after the sterilization, aseptic filter membrane is placed on the stainless steel filter membrane supporting pad, draw to separate liquid sample behind the homogeneous, pour in the stainless steel funnel to corresponding scale marks, open respective valves and carry out suction filtration.
In the present embodiment, can adopt the multiple film filtration unit, as long as can reach the effect of aseptic enriched microorganism.Fig. 2 adopts the microbial film filtration unit among the embodiment preferably, comprise stainless steel stent 21, at least one stainless steel filter, stainless-steel vacuum table 22, vacuum filtration bottle 23, vacuum pump 24, wherein vacuum meter 22 and stainless steel filter 23 are installed on the stainless steel stent 21, between stainless steel stent 21 and the vacuum pump 24 vacuum filtration bottle 25 is housed, stainless steel stent 21 is connected with the vacuum filtration bottle by pipe connecting 26 connections, and described filter 22 comprises stainless steel base 222 and the stainless steel funnel 221 that is installed on the stainless steel base.
The filtration unit of aforementioned micro organism membrane filter appts adopts all steel material, can select the multiple sterilization methods such as moist heat sterilization, dry sterilization, flame sterilization, especially can select flame sterilization, rapidly and efficiently, and speed of cooling is fast after the sterilization, can drop into rapidly to reuse.
In the preferred embodiment, described stainless steel funnel is connected on the stainless steel base by the mode of inlay card, concrete structure is: the stainless steel base is provided with at least three stainless steel projections, when the stainless steel funnel is subdued, projection is pressed into rear ejection, with funnel card jail, the mode by this kind inlay card is fixed on the stainless steel funnel on the stainless steel base, is convenient to picking and placeing of one-handed performance filter membrane.
Preferably, the stainless steel base is provided with stainless steel filter membrane supporting pad, and this stainless steel filter membrane supporting pad is formed by the compacting of stainless steel particle, supports evenly, the microorganism that assurance is trapped distributes at the filter membrane surface uniform, is convenient to subsequently microorganism culturing, counting or grain size analysis etc.
D. detect the microorganism 14 after the enrichment, after suction filtration is finished, touch the stainless steel funnel to a side, then upwards take out the stainless steel funnel, take off aseptic filter membrane with the antimagnetic type tweezer after the sterilization, carry out corresponding detection.
Enrichment can reach finite concentration to microorganism through suction filtration, according to actual needs, selects to count, detect the operations such as analysis, also can proceed follow-up microorganism culturing.
E. follow-up cultivation detects 15, and the aseptic filter membrane after the suction filtration enrichment is placed the sterilized culture dish that contains substratum, cultivates by inspecting standard, again detects after the cultivation.
Although the present invention with preferred embodiment openly as above; but it is not to limit the present invention; any those skilled in the art without departing from the spirit and scope of the present invention; can utilize method and the technology contents of above-mentioned announcement that technical solution of the present invention is made possible change and modification; therefore; every content that does not break away from technical solution of the present invention; to any simple modification, equivalent variations and modification that above embodiment does, all belong to the protection of technical solution of the present invention according to technical spirit of the present invention.
Claims (9)
1. micro-microorganism detection method in the polymictic liquid sample is characterized in that, may further comprise the steps:
Get a certain amount of liquid sample to be detected, put into membrane separation unit, then membrane separation unit is placed slap type homogenizer;
Velocity of flapping and homogenizing time are set, separate the solid impurity in the testing sample, and so that the liquid sample in the membrane separation unit is full and uniform;
Each parts to the microbial film filtration unit, comprise that stainless steel funnel and funnel web member sterilize, then connecting pipeline is closed each valve on the stainless steel stent, opens vacuum pump, regulate vacuum tightness, tweezers with after the sterilization are placed on aseptic filter membrane on the stainless steel filter membrane supporting pad, liquid sample behind the absorption separation homogeneous, pour in the stainless steel funnel to corresponding scale marks, open respective valves and carry out suction filtration;
After suction filtration is finished, touch the stainless steel funnel to a side, then upwards take out the stainless steel funnel, take off aseptic filter membrane with the antimagnetic type tweezer after the sterilization, carry out corresponding detection;
Aseptic filter membrane after the suction filtration enrichment is placed the sterilized culture dish that contains substratum, cultivate by inspecting standard, again detect after the cultivation.
2. detection method according to claim 1 is characterized in that, to the liquid sample of small amount, quantitative sample and diluent is added in the membrane separation unit simultaneously.
3. method according to claim 1 is characterized in that, to each parts of microbial film filtration unit, comprises that stainless steel funnel and funnel web member carry out flame sterilization.
4. the proofing unit of micro-microorganism in the polymictic liquid sample comprises membrane separation unit, slap type homogenizer, and microbial film filtration unit.
5. device according to claim 4, it is characterized in that, described membrane separation unit comprises the bag that two sides form, the sealing of bag four limits, top is for can remove part, and the bottom is the bag main body, inside is provided with filter membrane, the base of described filter membrane and top are individually fixed in base and the top of bag, and dual-side is individually fixed on the two sides of bag, and the width of filter membrane is less than the width of bag side.
6. device according to claim 5 is characterized in that, the dual-side of described filter membrane is individually fixed in the mid-way of bag two sides.
7. device according to claim 4, it is characterized in that, described microbial film filtration unit comprises stainless steel stent, at least one stainless steel filter, the stainless-steel vacuum table, the vacuum filtration bottle, vacuum pump, wherein vacuum meter and stainless steel filter are installed on the stainless steel stent by web member, the vacuum filtration bottle is housed between stainless steel stent and the vacuum pump, stainless steel stent is connected pipe connecting with the vacuum filtration bottle and connects, and described stainless steel filter comprises stainless steel base and the stainless steel funnel that is installed on the stainless steel base.
8. want 7 described devices according to right, it is characterized in that, described stainless steel funnel is connected on the stainless steel base by the mode of inlay card.
9. want 7 described devices according to right, it is characterized in that, described stainless steel base is provided with stainless steel filter membrane supporting pad.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104914100A (en) * | 2015-06-04 | 2015-09-16 | 苏州大学张家港工业技术研究院 | Site rapid bacterium visual detection method and site rapid bacterium visual detection kit |
| CN105296340A (en) * | 2014-09-19 | 2016-02-03 | 济南思拓新源医药科技有限公司 | Device for extracting some components in intestinal contents |
| CN106215499A (en) * | 2016-07-19 | 2016-12-14 | 浙江海洋大学 | A kind of zooplankton classified filtering device and application thereof |
| CN108827961A (en) * | 2018-04-25 | 2018-11-16 | 清远华新达饮品有限公司 | A kind of method of microorganism in measurement pure water |
| CN108872520A (en) * | 2018-08-14 | 2018-11-23 | 杭州绿洁水务科技股份有限公司 | A kind of water-quality test method, system and equipment |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105296340A (en) * | 2014-09-19 | 2016-02-03 | 济南思拓新源医药科技有限公司 | Device for extracting some components in intestinal contents |
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| CN104914100A (en) * | 2015-06-04 | 2015-09-16 | 苏州大学张家港工业技术研究院 | Site rapid bacterium visual detection method and site rapid bacterium visual detection kit |
| CN106215499A (en) * | 2016-07-19 | 2016-12-14 | 浙江海洋大学 | A kind of zooplankton classified filtering device and application thereof |
| CN106215499B (en) * | 2016-07-19 | 2018-11-23 | 浙江海洋大学 | A kind of zooplankter classified filtering device and its application |
| CN108827961A (en) * | 2018-04-25 | 2018-11-16 | 清远华新达饮品有限公司 | A kind of method of microorganism in measurement pure water |
| CN108872520A (en) * | 2018-08-14 | 2018-11-23 | 杭州绿洁水务科技股份有限公司 | A kind of water-quality test method, system and equipment |
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| CN103045713B (en) | 2015-05-27 |
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Effective date of registration: 20190520 Address after: 310000 Room 704, Building 4, Xigang Development Center, Xihu District, Hangzhou City, Zhejiang Province Patentee after: Hangzhou Prospective Biotechnology Co., Ltd. Address before: 310013 Unit 708, Block B, Zhejiang University Science Park, Hangzhou, Zhejiang Province Patentee before: Zhang Fan |